LOCUS       sequence-408950-        6646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6646)
  AUTHORS   Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, 
            Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser 
            SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC
  TITLE     A Human IgSF Cell-Surface Interactome Reveals a Complex Network of 
            Protein-Protein Interactions.
  JOURNAL   Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 
            10.1016/j.cell.2020.07.025.
  PUBMED    32822567
REFERENCE   2  (bases 1 to 6646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20- 
            20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
COMMENT     Sequence Label: pD649-HAsp-VSTM2L-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2222..2266
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2267..2284
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2424..2441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2430..2541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2659..2679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2664..2993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2844..2979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2906..2925
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3041..3088
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3107..3706
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3107..3126)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3603..3623
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3836..3957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3873..3892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3927..3946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4010..4801,4802..4870))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4633..4652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4871..4962)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5004..5023
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5029..5048
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5134..5722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5623..5642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5876..5893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5991..6008
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5992..6008
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6464..6480
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg ccacaagacc tgctggacat gctccttggg acaatcacgt
     1021 ttccggacac gccctgttca ccgagacacc tcacgatatg acagccagaa ccggcgagga
     1081 tgtggaaatg gcctgtagct ttagaggcag cggcagccct agctacagcc tggaaattca
     1141 gtggtggtac gtgcggagcc acagagactg gacagataag caggcctggg ccagcaatca
     1201 gctgaaagcc tctcagcaag aggacgccgg caaagaggcc acaaagatct ctgtcgtgaa
     1261 ggtcgtgggc agcaacatca gccacaagct gagactgagc agagtgaagc ctaccgacga
     1321 gggcacctac gagtgcagag tgatcgattt cagcgacggc aaggcccggc accacaaagt
     1381 gaaagcctat ctgagagtgc agcccggcga gaactctgtg ctgcatcttc ctgaagctcc
     1441 tccagctgct cctgctcctc ctccacctaa acctggcaaa gagctgcgga agagaagcgt
     1501 ggaccaagag gcctgctctc ttggcgcgcc gggcggcgac aagacccaca cctgcccccc
     1561 ctgcccagct ccagaactgc tgggagggcc ttccgtgttc ctgttccccc ccaagcccaa
     1621 ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg ccgtgtccca
     1681 cgaggaccca gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa
     1741 gaccaagccc agagaggagc agtacaacag cacctacagg gtggtgtccg tgctgaccgt
     1801 gctgcaccag gactggctga acggcaaaga atacaagtgc aaagtctcca acaaggccct
     1861 ggctgcccca atcgaaaaga caatcagcaa ggccaagggc cagccacggg agccccaggt
     1921 gtacaccctg ccccccagcc gggaggagat gaccaagaac caggtgtccc tgacctgtct
     1981 ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagagcaacg gccagcccga
     2041 gaacaactac aagaccaccc ccccagtgct ggacagcgac ggcagcttct tcctgtacag
     2101 caagctgacc gtggacaagt ccaggtggca gcagggcaac gtgttcagct gcagcgtgat
     2161 gcacgaggcc ctgcacaacc actacaccca gaagtccctg agcctgagcc ccggcggcag
     2221 cggtcttaat gatattttcg aggctcagaa gattgagtgg catgaacacc accaccatca
     2281 ccactaaggc cggcctgtag tcactggcac aacgcgttac ccggtaagcc aatcgggtat
     2341 acacggtcgt catactgcag acagggttct tctactttgc aagatagtct tgagtagtaa
     2401 aataaataga tagagaaaat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2461 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2521 ataaaatgag gaaattgcat cacaacactc aaccctatct cggtctattc ttttgattta
     2581 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2641 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2821 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2881 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     2941 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3001 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     3061 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga ccgagtacaa
     3121 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3181 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3241 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3301 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3361 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3421 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3481 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3541 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga catccgcgcc
     3601 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     3661 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg tgctacgaga
     3721 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3781 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     3841 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3901 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3961 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat taccaatgct
     4021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
     4141 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     4201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4321 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4861 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
     4981 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
     5041 accccttgct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     5101 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5161 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5221 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5281 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     5341 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5401 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5461 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5521 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5581 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5641 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5701 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5761 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5821 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5881 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     5941 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     6001 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     6061 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     6121 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     6181 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     6241 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     6301 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     6361 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     6421 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     6481 ttacgagctt catgcacagt ggcgttgact caatattggc cattagccat attattcatt
     6541 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     6601 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgtt
//
